Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes

Ullrich, Axel; Bell, Jeffrey R.; Chen, Ellson Y.; Herrera, Román; Petruzzelli, Lilli; Dull, Thomas J.; Gray, Andrew; Coussens, Lisa M.; Liao, Yu−Cheng; Tsubokawa, Makoto; Mason, Anthony J.; Seeburg, Peter H.; Grünfeld, Carl; Rosen, Ora M.; Ramachandran, J.

doi:10.1038/313756a0

Cited by 2,223 publications

(1,125 citation statements)

References 61 publications

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“…The human origin of the amplified products was confirmed by hybridization with 32 P-labeled probe (5Ј-CCT TGA GGT GAA CCA TCT CGA AGA TGA-3Ј) complementary to human INS-R mRNA nts 409-436. 17 The method was found to have a detection sensitivity of ෂ1 human cell in ෂ10 4 murine cells.…”

Section: Colony Typementioning

confidence: 99%

“…The presence of human DNA in murine tissue was detected using PCR primers designed to amplify sequence within the first exon of the human insulin receptor (INS-R) gene. 17 The PCR strategy employed was novel in that primers were designed to be non-hybridizable with murine INS-R sequence at their 3Ј end, thereby precluding amplification of murine DNA. The upstream primer was designed to hybridize at nts 162-183 and had the following sequence: 5Ј-GAT ATC CGG AAC AAC CTC ACT-3Ј.…”

Section: Colony Typementioning

confidence: 99%

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CD34+, kit+, rhodamine123low phenotype identifies a marrow cell population highly enriched for human hematopoietic stem cells

et al. 1998

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We hypothesized that human hematopoietic cells displaying a CD34 + , kit + , rhodamine123 low phenotype would be highly enriched for cells with stem-like properties. To test this hypothesis, we employed fluorescence activated cell sorting (FACS) to isolate cells with this phenotype from normal light density marrow mononuclear cells (MNC). CD34 + , kit + , rhodamine123 low cells comprised from 0.05-0.01% of the total MNC population. They were small, had scant cytoplasm, and contained nuclei with dense, hyperchromatic chromatin and inconspicuous nucleoli. Additional immunophenotyping revealed that these cells were CD33 − , CD38 − , CD20 − , and glycophorin A − . When plated in semisolid cultures containing optimal concentrations of IL-3, GM-CSF, KL, EPO, IL-6, and IL-1 these cells did not form colonies. However, when cultured over irradiated stromal cells, cobblestone areas were observed to form after 3 weeks, and harvested cells were able to initiate long-term cultures. To further demonstrate that these cells were indeed stem like, we also tested their ability to engraft and mature in immunocompromised (SCID) mice. Irradiated (400 cGy) SCID mice were transplanted with 2 × 10 3 candidate stem cells which were then injected with recombinant human growth factors every other day. Two months post-transplant the animals were sacrificed. PCR and FACS analysis of marrow and spleen cell samples revealed the presence of cells expressing human CD45 consistent with engraftment of human stem cells and the establishment of murine-human chimerism. Moreover, MNC isolated from transplanted mice formed unambiguously human BFU-E, CFU-GM and B cell colonies when stimulated with the appropriate growth factors. Accordingly, we have identified a relatively rapid and simple mechanism for isolating primitive human hematopoietic cells with stem cell-like properties. We anticipate that this strategy will be useful for experimental and therapeutic applications that require human stem cells in quantity.

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Section: Colony Typementioning

confidence: 99%

Section: Colony Typementioning

confidence: 99%

CD34+, kit+, rhodamine123low phenotype identifies a marrow cell population highly enriched for human hematopoietic stem cells

et al. 1998

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“…The insulin-like growth factor I (IGF-I) receptor is a member of the tyrosine kinase receptor family [1][2][3] and is composed of two α and two β subunits. The α subunits are entirely extracellular and bind IGF-I and IGF-II with high affinity.…”

Section: Introductionmentioning

confidence: 99%

“…Tyrosine phosAbbreviations used : CT, C-terminal ; EGF, epidermal growth factor ; GST, glutathione S-transferase ; IGF, insulin-like growth factor ; JCT, juxtamembrane ; MAP, mitogen-activated protein ; NGF, nerve growth factor ; PLC, phospholipase C ; WGA, wheat germ agglutinin. 1 Present address : Department of Pediatrics, Oregon Health Sciences University, Portland, OR 97201, U.S.A. 2 To whom correspondence should be addressed.…”

Section: Introductionmentioning

confidence: 99%

Interaction in vitro of the product of the c-Crk-II proto-oncogene with the insulin-like growth factor I receptor

Koval

Blakesley

Roberts

et al. 1998

Biochemical Journal

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The Crk proto-oncogene product is an SH2 and SH3 domaincontaining adaptor protein. We have previously demonstrated that Crk-II becomes rapidly tyrosine-phosphorylated in response to stimulation with insulin-like growth factor I (IGF-I) and might be involved in the IGF-I receptor signalling pathway. To determine whether this involvement includes the direct interaction of Crk-II with the cytoplasmic region of the receptor, studies were performed in itro with glutathione S-transferase (GST) fusion proteins containing various domains of Crk-II. The kinase assay in itro showed that activated IGF-I receptors efficiently phosphorylated the GST-Crk-II fusion protein. This phosphorylation was dependent on the presence of the SH2 domain and Tyr-221 located in the spacer region between the two SH3 domains. Mutation of Tyr-221 not only prevented phosphorylation of GST-Crk in itro, but also significantly increased the ability of GST-Crk proteins to co-precipitate activated IGF-I receptors from total cell lysates. Additional binding experiments in itro showed that Crk-II might interact with the phosphorylated IGF-I receptor through its SH2 domain. To elucidate which

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“…The insulin receptor structure has been elucidated through affinity labelling of receptors by chemical cross-linking with 125I-insulin, biosynthetic or cell-surface labelling of receptor subunits (for reviews see Jacobs&Cuatrecasas, 1981 ;Czech, 1984;VanObberghen, 1984). Recently, the complete amino-acid sequence of the human insulin receptor precursor was unravelled by application of recombinant DNA technology (Ullrich et al, 1985;Ebina et al, 1985). In spite ofthis progress the molecular mechanism of insulin action is still poorly comprehended regarding the events following the receptor binding and leading to the ultimate cellular responses.…”

Section: Introductionmentioning

confidence: 99%

Protein kinase activity of the insulin receptor

Gammeltoft¹,

Obberghen²

1986

Biochemical Journal

139

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The insulin receptor is an integral membrane glycoprotein (Mr approximately 300,000) composed of two alpha-subunits (Mr approximately 130,000) and two beta-subunits (Mr approximately 95,000) linked by disulphide bonds. This oligomeric structure divides the receptor into two functional domains such that alpha-subunits bind insulin and beta-subunits possess tyrosine kinase activity. The amino acid sequence deduced from cDNA of the single polypeptide chain precursor of human placental insulin receptor revealed that alpha- and beta-subunits consist of 735 and 620 residues, respectively. The alpha-subunit is hydrophilic, disulphide-bonded, glycosylated and probably extracellular. The beta-subunit consists of a short extracellular region which links the alpha-subunit through disulphide bridges, a hydrophobic transmembrane region and a longer cytoplasmic region which is structurally homologous with other tyrosine kinases like the src oncogene product and EGF receptor kinases. The cellular function of insulin receptors is dual: transmembrane signalling and endocytosis of hormone. The binding of insulin to its receptor on the cell membrane induces transfer of signal from extracellular to cytoplasmic receptor domains leading to activation of cell metabolism and growth. In addition, hormone-receptor complexes are internalized leading to intracellular proteolysis of insulin, whereas receptors are recycled to the membrane. These phenomena are kinetically well-characterized, but their molecular mechanisms remain obscure. Insulin receptor in different tissues and animal species are homologous in their structure and function, but show also significant differences regarding size of alpha-subunits, binding kinetics, insulin specificity and receptor-mediated degradation. We suggest that this heterogeneity of receptors may be linked to the diversity in insulin effects on metabolism and growth in various cell types. The purified insulin receptor phosphorylates its own beta-subunit and exogenous protein and peptide substrates on tyrosine residues, a reaction which is insulin-sensitive, Mn2+-dependent and specific for ATP. Tyrosine phosphorylation of the beta-subunit activates receptor kinase activity, and dephosphorylation with alkaline phosphatase deactivates the kinase. In intact cells or impure receptor preparations, a serine kinase is also activated by insulin. The cellular role of two kinase activities associated with the insulin receptor is not known, but we propose that the tyrosine- and serine-specific kinases mediate insulin actions on metabolism and growth either through dual-signalling or sequential pathways.(ABSTRACT TRUNCATED AT 400 WORDS)

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Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes

Cited by 2,223 publications

References 61 publications

CD34+, kit+, rhodamine123low phenotype identifies a marrow cell population highly enriched for human hematopoietic stem cells

CD34+, kit+, rhodamine123low phenotype identifies a marrow cell population highly enriched for human hematopoietic stem cells

Interaction in vitro of the product of the c-Crk-II proto-oncogene with the insulin-like growth factor I receptor

Protein kinase activity of the insulin receptor

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